Method for inhibiting microbial growth in liquid nutrients

ABSTRACT

A method for making a web having a metal-ion sequestering agent and treating a liquid using the web. The method of making the web includes electrostatically providing at least one metal-ion sequestering agent and/or antimicrobial agent. The web can be used to tread a liquid by placing the web into a tank and subjecting the liquid to the web.

CROSS REFERENCE TO RELATED APPLICATIONS

This is a continuation-in-part of U.S. patent application Ser. No.10/936,929 filed Sep. 9, 2004 which is a continuation-in-part of U.S.patent application Ser. No. 10/823,446 filed Apr. 13, 2004, U.S. Pat.No. 7,258,786.

Reference is also made to commonly assigned pending U.S. patentapplication Ser. No. 10/985,393 filed Nov. 10, 2004 entitled CONTAINERFOR INHIBITING MICROBIAL GROWTH IN LIQUID NUTRIENTS by David L. Patton,Joseph F. Bringley, Richard W. Wien, John M. Pochan, Yannick J. F. Leratand Narashimharao Dontula; and pending U.S. patent application Ser. No.10/985,377 filed Nov. 10, 2004 entitled CONTAINER FOR INHIBITINGMICROBIAL GROWTH IN LIQUID NUTRIENTS by David L. Patton, Joseph F.Bringley, Richard W. Wien, John M. Pochan, Yannick J. F. Lerat andWilliam J. Harrison the disclosures of which are incorporated herein byreference.

FIELD OF THE INVENTION

The present invention relates to a fluid container having a metal-ionsequestering agent for removing bio-essential metal ions from a liquidnutrient for inhibiting growth of microbes in the liquid nutrient.

BACKGROUND OF THE INVENTION

It has been recognized that small concentrations of metal ions play animportant role in biological processes. For example, Mn, Mg, Fe, Ca, Zn,Cu and Al are essential bio-metals, and are required for most, if notall, living systems. Metal ions play a crucial role in oxygen transportin living systems, and regulate the function of genes and replication inmany cellular systems. Calcium is an important structural element in thelife of bacteria regulating enzyme activity. Mn, Cu and Fe are involvedin metabolism and enzymatic processes. At high concentrations, metalsmay become toxic to living systems and the organism may experiencedisease or illness if the level cannot be controlled. As a result, theavailability, and concentrations, of metal ions in biologicalenvironments is a major factor in determining the abundance, growth-rateand health of plant, animal and micro-organism populations.

It has also been recognized that iron is an essential biologicalelement, and that all living organisms require iron for survival andreplication. Although, the occurrence and concentration of iron isrelatively high on the earth's surface, the availability of “free” ironis severely limited by the extreme insolubility of iron in aqueousenvironments. As a result, many organisms have developed complex methodsof procuring “free” iron for survival and replication.

Articles, such as food and beverage containers are needed that are ableto improve food quality, to increase shelf-life, to protect frommicrobial contamination, and to do so in a manner that is safe for theuser of such items and that is environmentally clean while providing forthe general safety and health of the public. Materials and methods areneeded to prepare articles having antimicrobial properties that areless, or not, susceptible to microbial resistance. Methods are neededthat are able to target and remove specific, biologically important,metal ions while leaving intact the concentrations of beneficial metalions.

During the process of filling containers with certain beverages andfoods, air borne pathogens enter the containers after the flashpasteurization or pasteurization part of the process. These pathogenssuch as yeast, spores, bacteria, etc. will grow in the nutrient richbeverage or food, ruining the taste or even causing hazardousmicrobiological contamination. While some beverages are packaged byaseptic means or by utilizing preservatives, many other beverages, forexample fruit juices, teas and isotonic drinks are “hot-filled”.“Hot-filling” involves the filling of a container with a liquid beveragehaving some elevated temperature (typically, at about 180-200° F.). Thecontainer is capped and allowed to cool, producing a partial vacuumtherein. The process of hot filling of beverages and foods is used tokill the pathogens, which enter the container during the filling of thebeverage or food containers. Hot filling requires containers be made ofcertain materials or constructed in a certain fashion such as thickerwalls to withstand the hot filling process. The energy required for hotfilling adds to the cost of the filling process. Temperatures requiredfor hot filling have a detrimental effect on the flavor of the beverage.Other methods of filling such as aseptic filling require large capitalexpenditures and maintaining class 5 clean room conditions.

U.S. Pat. No. 5,854,303 discloses a polymeric material incorporating apolyvalent cation chelating agent in an amount effective to inhibit thegrowth of a protozoan on the surface of contact lenses and in other eyecare products.

PROBLEM TO BE SOLVED BY THE INVENTION

The present invention is directed to the problem of the growth ofmicro-organism in liquids provided in containers that adversely affectsfood quality, shelf-life, to protect from microbial contamination, andto do so in a manner that is safe for the user of such and provides adegree of freedom in removing more than one containment in an efficientmanner.

SUMMARY OF THE INVENTION

In accordance with one aspect of the present invention, there isprovided a method for making a web having a metal-ion sequesteringagent, comprising the steps of:

a. providing a web; and

b. electrostatically providing at least one metal-ion sequestering agentand/or antimicrobial agent on the web.

In accordance with another aspect of the present invention there isprovided a method of treating a liquid comprising, the steps of:

a. providing a tank for containing a liquid to be treated;

b. providing a web in the tank, said web having said at least onemetal-ion sequestering agent and/or antimicrobial agent; and

c. subjecting a liquid provided in said web.

These and other aspects, objects, features and advantages of the presentinvention will be more clearly understood and appreciated from a reviewof the following detailed description of the preferred embodiments andappended claims and by reference to the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

In the detailed description of the preferred embodiments of theinvention presented below, reference is made to the accompanyingdrawings in which:

FIG. 1 illustrates a cross section of a fluid container made inaccordance with the prior art;

FIG. 2 is an enlarged partial cross sectional view of a portion of thecontainer of FIG. 1 illustrating a “free” iron ion sequestering agent;

FIG. 3 is a view similar to FIG. 2 illustrating a container made inaccordance with the present invention;

FIG. 4 illustrates a bottle with a bottle cap also made in accordancewith the present invention;

FIG. 5 is a schematic top plan view of the bottle and cap of FIG. 4;

FIG. 6 is an enlarged partial cross sectional view of the bottle and captaken along line 6-6 of FIG. 5;

FIG. 7 is a schematic view of a projecting member extending from amodified cap of FIG. 5 also made in accordance with the presentinvention;

FIG. 8 is an enlarged cross sectional view of the projecting member ofFIG. 7 as taken along line 8-8;

FIG. 9 is a schematic view of another embodiment of the presentinvention illustrating one method for applying a coating to the interiorsurface of a bottle made in accordance with the present invention;

FIG. 10 is an enlarged partial cross sectional view of a portion of thebottle of FIG. 9 illustrating the sprayed coating of the ionsequestering agent;

FIG. 11 is a schematic view of another fluid container made inaccordance with the present invention such as a juice box;

FIG. 12 is an enlarged partial cross sectional view of the juice boxtaken along line 12-12 of FIG. 11;

FIG. 13 is a schematic view of yet another fluid container such as astand up pouch made in accordance with the present invention;

FIG. 14 is an enlarged partial cross sectional view of the stand uppouch taken along line 14-14 of FIG. 13;

FIG. 15 is a schematic view of still another embodiment of a fluidcontainer such as a bag also made in accordance with the presentinvention;

FIG. 16 is an enlarged partial cross sectional view of a portion of thebag of FIG. 15 as indicated by circle 16;

FIG. 17 is a cross-sectional view of a web that can be used in themanufacture of a box, pouch or bag showing a coating assembly forcoating a hydrophilic layer containing a metal-ion sequestering agent;

FIG. 18 is a schematic view of yet another fluid container, such as acan, made in accordance with the present invention;

FIG. 19 is a cross sectional view of FIG. 18 as taken along line 19-19;

FIG. 20 is a cross sectional view of a filter assembly made inaccordance with the present invention;

FIG. 21 is a cross sectional view of a fluid bed ion exchange assemblymade in accordance with the present invention;

FIG. 22 is an enlarged partial view of a portion of the fluid bed ionexchange assembly of FIG. 21 as identified by circle 22 illustrating ametal-ion sequestering agent;

FIG. 23 is a schematic view of electro-photographic device for applyinga pattern of metal ion chelating agents to a web, such as is used tomake a box, pouch or bag in accordance with the present invention;

FIG. 24 is a schematic top view of the web and device of FIG. 23;

FIGS. 25 and 26 are both schematics of a holding tank with a webassembly;

FIG. 27 is a schematic diagram of yet another embodiment of a holdingtank with a web also made in accordance with the present invention; and

FIG. 28 is a schematic diagram of yet still another embodiment of aholding tank with a web also made in accordance with the presentinvention.

FIG. 29 is a bar graph illustrating fungostatic and fungicidal effectsof the invention on example yeast populations.

DETAILED DESCRIPTION OF THE INVENTION

The growth of microbes in an article such as a fluid containercontaining a liquid nutrient comprising a liquid nutrient can beinhibited by placing metal-ion sequestering agents, as described in U.S.patent application Ser. No. 10/822,940 filed Apr. 13, 2004 entitledDERIVATIZED NANOPARTICLE COMPRISING METAL-ION SEQUESTRANT by Joseph F.Bringley, and U.S. patent application Ser. No. 10/822,929 filed Apr. 13,2004 entitled COMPOSITION OF MATTER COMPRISING POLYMER AND DERIVATIZEDNANOPARTICLES by Joseph F. Bringley et al. capable of removing adesignated metal ion for example, Mn, Fe, Ca, Zn, Cu and Al from saidliquid nutrients, in contact with the nutrient. Intimate contact isachieved by incorporating the metal-ion sequestering agent as anintegral part of the support structure of the article. For example, onecan control the concentration of “free” iron in the liquid nutrient heldby the article by placing an iron sequestering agent in the walls of thecontainer, which in turn controls the growth rates, and abundance ofmicro-organisms. The articles of the invention further contain aneffective amount of an antimicrobial agent, which quickly reduces thepopulation of microbes to a manageable level, and insures theeffectiveness of metal-ion sequestering or binding agents. The invention“starves” the remaining micro-organisms of minute quantities ofessential nutrients (metal-ions) and hence limits their growth andreduces the risk due to bacterial, viral and other infectious diseases.The article, such as a container, may be used for holding a food orbeverage.

The term inhibition of microbial-growth, or a material which “inhibits”microbial growth, is used by the authors to mean materials which eitherprevent microbial growth, or subsequently kills microbes so that thepopulation is within acceptable limits, or materials which significantlyretard the growth processes of microbes or maintain the level ormicrobes to a prescribed level or range. The prescribed level may varywidely depending upon the microbe and its pathogenicity; generally it ispreferred that harmful organisms are present at no more than 10organisms/ml and preferably less than 1 organism/ml. Antimicrobialagents which kill microbes or substantially reduce the population ofmicrobes are often referred to as biocidal materials, while materialswhich simply slow or retard normal biological growth are referred to asbiostatic materials. The preferred impact upon the microbial populationmay vary widely depending upon the application, for pathogenic organisms(such as E. coli O157:H7) a biocidal effect is more preferred, while forless harmful organisms a biostatic impact may be preferred. Generally,it is preferred that microbiological organisms remain at a level whichis not harmful to the consumer or user of that particular article.

Metal-ion sequestering agents may be incorporated into articles byplacing the metal-ion sequestering agents on the surface of the article,or by putting the metal-ion sequestering agents within the materialsused to form the article. In all instances, the metal-ion sequesteringagents must be capable of contacting the food or beverage held by thecontainer.

Referring to FIG. 1, there is illustrated a cross-sectional view of atypical prior art container. In the embodiment illustrated, thecontainer comprises a bottle 5 holding a liquid nutrient 10, for examplean isotonic liquid. Drinks such as Gatorade™ or PowerAde™ are examplesof isotonic drinks/liquids. The container 5 may be made of one or morelayers of a plastic polymer using various molding processes known bythose skilled in the art. Examples of polymers used in the manufactureof bottles are PET (polyethylene terephthalate), PP (polypropylene),LDPE (low density polyethylene) and HDPE (high density polyethylene).FIG. 2 illustrates a plastic bottle 5 formed using two differentpolymeric layers 15 and 20. However it is to be understood that thecontainer 5 may comprise any desired number of layers.

A fluid container made in accordance with the present invention isespecially useful for containing a liquid nutrient having a pH equal toor greater than about 2.5. The container is designed to have an interiorsurface having a metal-ion sequestering agent for removing a designatedmetal ion from a liquid nutrient for inhibiting growth of microbes insaid liquid nutrient. It is preferred that the metal-ion sequestrant isimmobilized within the materials forming the container or is immobilizedwithin a polymeric layer directly in contact with the beverage or liquidnutrient. It is further preferred that the metal-ion sequestering agentis immobilized on the surface(s) of said container. This is importantbecause metal-ion sequestrants that are not immobilized may diffusethrough the material or polymeric layers of the container and dissolveinto the contents of the beverage. Metal ions complexed by dissolvedsequestrants will not be sequestered within the surfaces of thecontainer but may be available for use by micro-organisms.

It is preferred that the sequestering agent is immobilized on thesurface(s) of said container and has a high-affinity for biologicallyimportant metal ions such as Mn, Mg, Zn, Cu and Fe. It is furtherpreferred that the immobilized sequestering agent has a high-selectivityfor biologically important metal ions such as Mn, Mg, Zn, Cu and Fe. Itis preferred that said sequestering agent has a high-selectively forcertain metal ions but a low-affinity for at least one other ion. It isfurther preferred that said certain metal ions comprises Mn, Mg, Zn, Cuand Fe and said other at least one ion comprises calcium. This ispreferred because some metal ions such as calcium, sodium and potassiummay be beneficial to the taste and quality of the food, and are usuallyvery highly abundant in foodstuffs and in liquid extrudates offoodstuffs. It is preferred that said metal-ion sequestering agent isimmobilized on the surface(s) of said container and has a stabilityconstant greater than 10¹⁰ with iron (III), more preferably greater than10²⁰ with iron (III), and most preferably greater than 10³⁰ with iron(III). This is preferred because iron is an essential nutrient forvirtually all micro-organisms, and sequestration of iron may mostbeneficially limit the growth of micro-organisms.

In a particularly preferred embodiment, the invention provides a fluidcontainer wherein said metal-ion sequestering agent comprisesderivatized nanoparticles comprising inorganic nanoparticles having anattached metal-ion sequestrant, wherein said inorganic nanoparticleshave an average particle size of less than 200 nm and the derivatizednanoparticles have a stability constant greater than 10¹⁰ with iron(III). It is preferred that the inorganic nanoparticles have an averageparticle size of less than 100 nm. It is preferred that said metal-ionsequestrant is attached to the nanoparticle by reacting the nanoparticlewith a silicon alkoxide intermediate of the sequestrant having thegeneral formula:Si(OR)_(4-x)R′_(x;)wherein x is an integer from 1 to 3;

-   R is an alkyl group; and-   R′ is an organic group containing an alpha amino carboxylate, a    hydroxamate, or a catechol. Derivatized nanoparticles useful for    practice of the invention are described in detail in U.S. patent    application Ser. No. 10/822,940 filed Apr. 13, 2004 entitled    DERIVATIZED NANOPARTICLE COMPRISING METAL-ION SEQUESTRANT by    Joseph F. Bringley.

In a preferred embodiment the metal-ion sequestering agent isimmobilized in a polymeric layer, and the polymeric layer contacts thefluid contained therein. The metal-ion sequestrant may be formedintegrally within the materials comprising the bottle or may becontained within a polymeric layer directly in contact with the beverageor liquid nutrient. It is preferred that the polymer is permeable towater. It is preferred that the metal-ion sequestering agent comprisesare 0.1 to 50.0% by weight of the polymer. Polymers useful for practiceof the invention are described in detail in U.S. patent application Ser.No. 10/823,453 filed Apr. 13, 2004 entitled ARTICLE FOR INHIBITINGMICROBIAL GROWTH by Joseph F. Bringley et al.

In a preferred embodiment, the metal-ion sequestering agent comprises analpha amino carboxylate, a hydroxamate, or a catechol functional group.Metal-ion sequestrants suitable for practice of the invention includeethylenediaminetetraacetic acid (EDTA), ethylenediaminetetraacetic aciddisodium salt, diethylenetriaminepentaacetic acid (DTPA),Hydroxylpropylenediaminetetraacetic acid (DPTA), nitrilotriacetic acid,triethylenetetraaminehexaacetic acid, N,N-bis(o-hydroxybenzyl)ethylenediamine-N,N diacteic acid, andethylenebis-N,N′-(2-o-hydroxyphenyl)glycine, acetohydroxamic acid, anddesferroxamine B (the iron chelating drug desferal), catechol,disulfocatechol, dimethyl-2,3-dihydroxybenzamide, mesitylenecatecholamide (MECAM) and derivatives thereof,1,8-dihydroxynaphthalene-3,6-sulfonic acid, and2,3-dihydroxynaphthalene-6-sulfonic acid, and siderophores moleculesnaturally synthesized by micro-organisms which have a very high affinityfor Fe. Metal-ion sequestering agents suitable for use in the inventionare described at length in U.S. patent application Ser. No. 10/822,940filed Apr. 13, 2004 entitled DERIVATIZED NANOPARTICLE COMPRISINGMETAL-ION SEQUESTRANT by Joseph F. Bringley et al.

The antimicrobial active material of antimicrobial agent may be selectedfrom a wide range of known antibiotics and antimicrobials. Anantimicrobial material may comprise an antimicrobial ion, moleculeand/or compound, metal ion exchange materials exchanged or loaded withantimicrobial ions, molecules and/or compounds, ion exchange polymersand/or ion exchange latexes, exchanged or loaded with antimicrobialions, molecules and/or compounds. Suitable materials are discussed in“Active Packaging of Food Applications” A. L. Brody, E. R. Strupinskyand L. R. Kline, Technomic Publishing Company, Inc. Pennsylvania (2001).Examples of antimicrobial agents suitable for practice of the inventioninclude benzoic acid, sorbic acid, nisin, thymol, allicin, peroxides,imazalil, triclosan, benomyl, metal-ion release agents, metal colloids,anhydrides, and organic quaternary ammonium salts. Preferredantimicrobial reagents are metal ion exchange reagents such as silversodium zirconium phosphate, silver zeolite, or silver ion exchange resinwhich are commercially available. The antimicrobial agent may beprovided in a layer 15 having a thickness “y” of between 0.1 microns and100 microns, preferably in the range of 1.0 and 25 microns.

In another preferred embodiment, the antimicrobial agent comprising acomposition of matter comprising an immobilized metal-ionsequestrant/antimicrobial comprising a metal-ion sequestrant that has ahigh stability constant for a target metal ion and that has attachedthereto an antimicrobial metal-ion, wherein the stability constant ofthe metal-ion sequestrant for the antimicrobial metal-ion is less thanthe stability constant of the metal-ion sequestrant for the targetmetal-ion. These are explained in detail in U.S. patent application Ser.No. 10/868,626 filed Jun. 15, 2004 entitled AN IRON SEQUESTERINGANTIMICROBIAL COMPOSITION by Joseph F. Bringley et al.

In a preferred embodiment, the antimicrobial agent comprising a metalion exchange material, which is exchanged with at least oneantimicrobial metal ion selected from silver, copper, gold, nickel, tinor zinc.

Referring to FIG. 3, there is illustrated a cross-section view of animproved fluid container such as shown in FIG. 1 made in accordance withthe present invention. The container, which in the embodimentillustrated is a bottle, is made of a material that comprises a barrierlayer 22, an outer polymeric layer 20 and an inner polymeric layer 40between said barrier layer 22 and outer polymeric layer 20. The innerpolymeric layer 40 contains a metal-ion sequestrant 35. The barrierlayer 22 preferably does not contain the metal-ion sequestrant 35. Theouter layer 20 may provide several functions including improving thephysical strength and toughness of the article and resistance toscratching, marring, cracking, etc. However, the primary purpose of thebarrier layer 22 is to provide a barrier through which micro-organisms25 present in the contained fluid cannot pass. It is important to limitor eliminate, in certain applications, the direct contact ofmicro-organisms 25 with the metal-ion sequestrant 35 or the layercontaining the metal-ion sequestrant 35, since many micro-organisms 25,under conditions of iron deficiency, may bio-synthesize molecules whichare strong chelators for iron and other metals. These bio-syntheticmolecules are called “siderophores” and their primary purpose is toprocure iron for the micro-organisms 25. Thus, if the micro-organisms 25are allowed to directly contact the metal-ion sequestrant 35, they mayfind a rich source of iron there and begin to colonize directly at thesesurfaces. The siderophores produced by the micro-organisms may competewith the metal-ion sequestrant for the iron (or other bio-essentialmetal) at their surfaces. However the energy required for the organismsto adapt their metabolism to synthesize these siderophores will impactsignificantly their growth rate. Thus, one object of the invention is tolower growth rate of organisms in the contained liquid. Since thebarrier layer 22 of the invention does not contain the metal-ionsequestrant 35, and because micro-organisms are large, themicro-organisms may not pass or diffuse through the barrier layer 22.The barrier layer 22 thus prevents contact of the micro-organisms withthe polymeric layer 40 containing the metal-ion sequestrant 35 of theinvention. It is preferred that the barrier layer 22 is permeable towater. It is preferred that the barrier layer 22 has a thickness “x” inthe range of 0.1 microns to 10.0 microns. It is preferred that microbesare unable to penetrate, to diffuse or pass through the barrier layer22. Sequestrant 35 with a sequestered metal ion is indicated by numeral35′.

Still referring again to FIG. 3, the enlarged sectioned view of thefluid container 5 shown in 3, illustrates a bottler having barrier layer22, which is in direct contact with the liquid nutrient 10, an innerpolymeric layer 40 and an outer polymeric layer 20. However, the bottleof FIG. 2 comprises an inner polymeric layer 15 that does not containany metal-ion sequestering agents. In the prior art bottle illustratedin FIG. 2, the micro-organisms 25 are free to gather the “free” ironions 30. In the example shown in FIG. 3, the inner polymer 40 containsan immobilized metal-ion sequestering agent 35 such as EDTA. In orderfor the metal-ion sequestering agent 35 to work properly, the innerpolymer 40 containing the metal-ion sequestering agent 35 must bepermeable to aqueous media. Preferred polymers for layers 22 and 40 ofthe invention are polyvinyl alcohol, cellophane, water-basedpolyurethanes, polyester, nylon, high nitrile resins,polyethylene-polyvinyl alcohol copolymer, polystyrene, ethyl cellulose,cellulose acetate, cellulose nitrate, aqueous latexes, polyacrylic acid,polystyrene sulfonate, polyamide, polymethacrylate, polyethyleneterephthalate, polystyrene, polyethylene, polypropylene orpolyacrylonitrile. A water permeable polymer permits water to movefreely through the polymer 40 allowing the “free” iron ion 30 to reachand be captured by the agent 35. An additional barrier 22 may be used toprevent the micro-organism 25 from reaching the inner polymer material40 containing the metal-ion sequestering agent 35. Like the innerpolymer material 40, the barrier layer 22 must be made of a waterpermeable polymer as previously described. The micro-organism 25 is toolarge to pass through the barrier 22 or the polymer 40 so it cannotreach the sequestered iron ion 30 now held by the metal-ion sequesteringagent 35. By using the metal-ion sequestering agents 35 to significantlyreduce the amount of “free” iron ions 30 in the liquid nutrient 10, thegrowth of the micro-organism 25 is eliminated or severely reduced.

In the embodiment shown in FIGS. 4, 5, and 6 the metal-ion sequesteringagent 35 is contained in the bottle cap 50 instead of on the insidesurface of the bottle 5. An inner portion 45 of the cap 50, which is inintimate contact with the liquid nutrient 10, is made of a hydrophilicpolymer 55 containing the metal-ion sequestering agent 35 such as EDTAas described above. In some situations, the bottle may need to be placedin the inverted position in order for the sequestrant to become incontact with the contained nutrient. The cap 50 may also have thebarrier layer 22 to further prevent the micro-organisms 25 from reachingthe sequestered “free” iron 30. In another embodiment (not shown) thecap sealing material could be an open cell foamed structure whose cellwalls are coated with the sequestering material.

In still another embodiment, the sequestering agent 35 may be in ahydrophilic polymeric insert 52 that is placed in the bottle 5 asillustrated in FIG. 4. The insert 52 may be instead of or in addition tothe sequestrant in the cap 50 or interior of the bottle. The insert 52is placed in the bottle 5 but unfolds making it too large to exit thebottle 5. In another version, the insert 52 is molded into the bottom ofthe bottle 5.

Referring to FIGS. 7 and 8, there is illustrated another modifiedembodiment of a container made in accordance with the present invention,like parts indicating like parts and operation as previously described.In this embodiment the metal-ion sequestering agent 35 is contained in aprojecting member 60 that extends from cap 50 into the bottle 5 so thatit will be in intimate contact with the liquid nutrient 10. In theembodiment, the projecting member is in the configuration of a strawthat can later be used to drink the liquid content in the bottle. Likethe hydrophilic polymer material lining of the inside of the bottle 5and bottle cap 50, the extension 60 or straw is made of a hydrophilicpolymer 65 containing the metal-ion sequestering agents 35 such as EDTAas described in FIG. 3. When the bottle 5 is filled with the liquidnutrient 10 such as an isotonic, and is capped, the straw 60 protrudesfrom the cap 50 into the solution 10 allowing the “free” iron ions 35 tobe sequestered from the liquid nutrient 10. The straw 60 may also havethe barrier layer 22 to further prevent the micro-organisms 25 fromreaching the sequestered “free” iron ions 30. The outer layer 20 mayalso be made of a material similar to barrier layer 22 so that “free”iron ions 30 can reach the sequestrant 35 from the outside of the straw60.

In the example shown the extension is a straw but the extension can beof any shape just as long as it extends into the food or beverageestablishing intimate contact.

Referring to FIGS. 9 and 10, there is illustrated another embodiment ofa bottle 5 made in accordance with the present invention. In thisembodiment, the metal-ion sequestering agent 35 is applied to the insidesurface 80 of the bottle 5 by spraying a metal-ion sequestering agent35, for example EDTA, on to the inside surface of the bottle through asupply tube 85 using a spherical shaped nozzle assembly 90. The nozzleassembly 90 is moved up and down in the direction of the arrow 95 whilethe metal-ion sequestering agent 35 is sprayed as indicated by thearrows 100. The method of applying coatings to glass, metal or plasticcontainers is well known to those skilled in the art. FIG. 10illustrates an enlarged partial cross sectional view of the portion ofthe bottle of FIG. 9 where the spray coating 105 of the ion sequesteringagent 35 has been applied. As previously discussed in FIG. 3, likenumerals indicate like parts and operations. It is of course understoodthat the inner layer containing the sequestrant may be applied or formedon the inside surface of the container in any appropriate manner. Thebottle 5 in this embodiment may be made of any appropriate plastic orglass material.

Referring to FIGS. 11 and 12, there is illustrated yet another modifiedcontainer 110 made in accordance with the present invention. Inparticular the container comprises juice/drink box 110 for containing aliquid beverage. The box 110 is made of a sheet material that comprisesinner layer 115, a middle layer 120 made of a hydrophobic polymermaterial, and an outer layer 125. The inner layer 115 is in directcontact with the liquid nutrient 10 and is made of a hydrophilic polymercontaining the metal-ion sequestering agent 35 such as EDTA as describedabove in FIG. 3. As previously discussed in FIG. 3, like numeralsindicate like parts and operations. The outer layer 125 may comprise afoil wrap.

Referring to FIGS. 13 and 14, there is illustrated yet another modifiedembodiment of a container 130 made in accordance with the presentinvention. In the embodiment, the container comprises a stand up pouch130. The pouch 130 comprises an inner layer 135 made of a hydrophilicpolymer material, and an outer layer 140. The outer layer 140 may bemade of a polymer such as Mylar™ with a metalized coating 145. The innerlayer 135 is in direct contact with the liquid nutrient 10 and is madeof a hydrophilic polymer containing the metal-ion sequestering agent 35such as EDTA as described above in FIG. 3. The stand up pouch 130 mayalso have the barrier layer 22 not shown to further prevent themicro-organisms 25 from reaching the sequestered “free” iron 30. Aspreviously discussed in FIG. 3, like numerals indicate like parts andoperations.

Referring to FIGS. 15 and 16, there is illustrated still anothermodified container made in accordance with the present invention. Inthis embodiment the container comprises a bag 150. The bag 150, which isintended to hold an aqueous material, comprises an inner layer 155 madeof a hydrophobic polymer material and an outer layer 160. The outerlayer 140 may be made of a polymer such as polyethylene terephthalate.The inner layer 155 is in direct contact with the aqueous material 155and is made of a hydrophilic polymer containing the metal-ionsequestering agent 35 such as EDTA as described above in FIG. 3. The bag150 may also have the barrier layer 22 not shown to further prevent themicro-organisms 25 from reaching the sequestered “free” iron 30. Aspreviously discussed in FIG. 3, like numerals indicate like parts andoperations.

The juice box 110, the pouch 130 and the bag 150 may be constructed froma base web 170 as illustrated in FIG. 17. After the base web 170 isformed, the hydrophilic layer 175 is applied via a coating assembly 180comprised of a reservoir 185, an applicator 190 and a drive mechanism(not shown) to form the hydrophilic inner layer 175 containing themetal-ion sequestering agent 35 as described above in FIG. 3. Othermethods of forming and of making webs and applying a coating such ascoextrusion may be used. It is of course understood that any suitabletechnique or process may be used for applying a coating on supportingweb as long as the coating has the appropriate sequestrant.

Referring to FIGS. 18 and 19 there is illustrated and modified container220 made in accordance with the present invention. In this embodiment,the container 220 comprises a can. The can 200 is made of a metalmaterial such as aluminum or steel, and has a top and a bottom, whichmay or may not be made as separate piece. The can 200 may also have alining 205, which is in direct contact with the aqueous material 155 andintended to prevent corrosion of the metal by the contents of the can.The construction of metal cans is well known by one skilled in the art.The lining 205 may include a hydrophilic polymer containing themetal-ion sequestering agent 35 or have a hydrophilic polymer strip 210containing metal-ion sequestering agent 35 made as part of lining 205 ofthe can 200. The strip 210 may have a width “w” of between 1 millimeterand 30 millimeters and be spaced at intervals around the insidecircumference of the can 200 and a depth “d” of −1.0 to 10 micrometers.In still another embodiment, the sequestering agent 35 may be in ahydrophilic polymeric insert 52. The insert 52 is placed in the can 200but unfolds making it too large to exit the can 200. The insert 52 maybe simply placed on the bottom of the container or if desired secured tothe interior surface of the container in some fashion. The metal-ionsequestering agent performs as previously described above in FIG. 3.

Referring to FIG. 20, there is illustrated a cross-sectional view of afilter assembly 220 comprising an inlet port 225, an outlet port 230,and a filter 235. The filter 235 contains an immobilized metal-ionsequestering agent as previously described. As the solution flowsthrough the filter assembly 220 in the direction indicated by the arrows240, and through the filter 235 the metal ions in the solution aresequestered and removed by the metal-ion sequestering agent 245.

Referring to FIG. 21, there is illustrated a cross sectional view of afluid bed ion exchange assembly 250 comprising a holding tank 255, aninlet port 260, an outlet port 265, and a fluid bed 270 containing ametal-ion sequestering material 275 made in accordance with the presentinvention. The solution 280 flows into the fluid bed ion exchangeassembly 250 via inlet port 260 as indicated by arrow 285 through themetal-ion sequestering material 275 in fluid bed 270 as indicated byarrows 290 and out the outlet port as indicated by arrow 295.

FIG. 22 is an enlarged partial view of a portion of the fluid bed 270containing a metal-ion sequestering material 275. An example of themetal-ion sequestering material 275 comprises a core material 300 and ashell material 305 made of the metal-ion sequestering agent 35 asdescribed in U.S. patent application Ser. No. 10/822,940 filed Apr. 13,2004 entitled DERIVATIZED NANOPARTICLE COMPRISING METAL-ION SEQUESTRANTby Joseph F. Bringley et al. As previously described above in FIG. 21,the solution 280 containing “free” metal ions 310 flows through thefluid bed 270 as indicated by the arrows 315. As the solution 280 flowsthrough the fluid bed 270 the shell material 305 made of the metal-ionsequestering agent 35 gathers the metal ions 320 removing them from thesolution, which then flow out through the outlet port 265.

While in the embodiments discussed, the iron sequestering agent orantimicrobial agent may be provided only on a portion of the contactingsurface of the bottle or other container. For example, but not limitedto, the agents may be provided only on the body portion of a bottle andnot the neck portion.

While in many of the embodiments illustrated a barrier layer is notdiscussed, it is to be understood that a barrier layer 22 may beprovided in any of the embodiments for preventing the microbes(micro-organism) from contacting the sequestrant.

EXAMPLES AND COMPARISON EXAMPLES

Materials:

Colloidal dispersions of silica particles were obtained from ONDEO NalcoChemical Company. NALCO® 1130 had a median particle size of 8 nm, a pHof 10.0, a specific gravity of 1.21 g/ml, a surface area of about 375m²/g, and a solids content of 30 weight percent.N-(trimethoxysilylpropyl ethylenediamine triacetic acid, trisodium saltwas purchased from Gelest Inc., 45% by weight in water.

Preparation of derivatized nanoparticles. To 600.00 g of silica NALCO®1130 (30% solids) was added 400.00 g of distilled water and the contentsmixed thoroughly using a mechanical mixer. To this suspension, was added49.4 g of N-(trimethoxysilyl) propylethylenediamine triacetic acid,trisodium salt in 49.4 g distilled water with constant stirring at arate of 5.00 ml/min. At the end of the addition the pH was adjusted to7.1 with the slow addition of 13.8 g of concentrated nitric acid, andthe contents stirred for an hour at room temperature. Particle sizeanalysis indicated an average particle size of 15 nm. The percent solidsof the final dispersion was 18.0%.Preparation of the immobilized metal-ion sequestrant/antimicrobial:200.0 g of the above derivatized nanoparticles were washed withdistilled water via dialysis using a 6,000-8,000 molecular weight cutofffilter. The final ionic strength of the solution was less than 0.1millisemens. To the washed suspension was then added with stirring 4.54ml of 1.5 M AgNO₃ solution, to form the immobilized metal-ionsequestrant/antimicrobial.Preparation of Polymeric Layers of Immobilized Metal-Ion Sequestrantsand Sequestrant/Antimicrobials.

Coating 1 (comparison). A coating solution was prepared as follows: 8.8g of a 40% solution of the polyurethane Permax 220 (Noveon Chemicals)was combined with to 90.2 grams of pure distilled water and 1.0 g of a10% solution of the surfactant OLIN 10G was added as a coating aid. Themixture was then stirred and blade-coated onto a polymeric support usinga 150 micron doctor blade. The coating was then dried at 40-50° C., toproduce a film having 5.4 g/m² of polyurethane.

Coating 2. A coating solution was prepared as follows: 171.2 grams ofthe derivatized nanoparticles prepared as described above were combinedwith 64.8 grams of pure distilled water and 62.5 g of a 40% solution ofthe polyurethane Permax 220 (Noveon Chemicals). 1.5 g of a 10% solutionof the surfactant OLIN 10G was added as a coating aid. The mixture wasthen stirred and blade-coated onto a polymeric support using a 150micron doctor blade. The coating was then dried at 40-50° C., to producea film having 5.4 g/m² of the derivatized nanoparticles and 5.4 g/m² ofpolyurethane.Coating 3. A coating solution was prepared as follows: 171.2 grams ofthe derivatized nanoparticles prepared as described above were combinedwith 33.5 grams of pure distilled water and 93.8 g of a 40% solution ofthe polyurethane Permax 220 (Noveon Chemicals). 1.5 g of a 10% solutionof the surfactant OLIN 10G was added as a coating aid. The mixture wasthen stirred and blade-coated onto a polymeric support using a 150micron doctor blade. The coating was then dried at 40-50° C., to producea film having 5.4 g/m² of the derivatized nanoparticles and 8.1 g/m² ofpolyurethane.Coating 4. A coating solution was prepared as follows: 138.9 grams ofthe derivatized nanoparticles prepared as described above were combinedwith 97.1 grams of pure distilled water and 62.5 g of a 40% solution ofthe polyurethane Permax 220 (Noveon Chemicals). 1.5 g of a 10% solutionof the surfactant OLIN 10G was added as a coating aid. The mixture wasthen stirred and blade-coated onto a polymeric support using a 150micron doctor blade. The coating was then dried at 40-50° C., to producea film having 4.4 g/m² of the derivatized nanoparticles and 5.4 g/m² ofpolyurethane.Coating 5. A coating solution was prepared as follows: 12.8 grams of theimmobilized metal-ion sequestrant/antimicrobial suspension prepared asdescribed above was combined with to 77.4 grams of pure distilled waterand 8.8 g of a 40% solution of the polyurethane Permax 220 (NoveonChemicals). 1.0 g of a 10% solution of the surfactant OLIN 10G was addedas a coating aid. The mixture was then stirred and blade-coated onto apolymeric support using a 150 micron doctor blade. The coating was thendried at 40-50° C., to produce a film having 2.7 g/m² of the immobilizedmetal-ion sequestrant/antimicrobial, 0.06 g/m² silver-ion and 5.4 g/m²of polyurethane.

Testing Methodology

A test similar to ASTM E 2108-01 was conducted where a piece of acoating of known surface area was contacted with a solution inoculatedwith micro-organisms. In particular a piece of coating 1×1 cm was dippedin 2 ml of growth medium (Trypcase Soy Agar 1/10), inoculated with 2000CFU of Candida albicans (ATCC-1023) per ml. Special attention was madeto all reagents to avoid iron contamination with the final solutionhaving an iron concentration of 80 ppb before contact with the coating.

Micro-organism numbers in the solution were measured daily by thestandard heterotrophic plate count method.

The bar graph shown in FIG. 29 demonstrates the effectiveness of theinventive examples. The yeast population which was exposed to thecomparison coating 1 (which contained no derivatized nanoparticles)showed a growth factor of one thousand during 48 hours (a 1000-foldincrease in population). The yeast population which was exposed to theexample coatings 2-4 (containing derivatized nanoparticles) showedgrowth factors of only 1-4. This is indicative of a fungostatic orbio-static effect in which the population of organisms is kept at aconstant or near constant level, even in the presence of a mediumcontaining adequate nutrient level. The yeast population which wasexposed to the example coating 5 (derivatized nanoparticles that hadbeen ion exchanged with silver ion—a known antimicrobial) showed afungicidal effect (the yeast were completely eliminated). The low levelof silver when coated by itself without the nanoparticles would not beexpected to exhibit this complete fungicidal effect, and there appearsto be a synergistic effect between the iron sequestration and therelease of antimicrobial silver.

As can be seen from the bar graph illusttrated in FIG. 29, significantimproved results may be obtained when a metal-ion sequestering agent isused in conjunction with an antimicrobial agent. The combined agentsreduced the level of microbes to lower level than when first introducedand then maintained the reduced level of microbes in the liquidnutrient.

Now referring to FIGS. 23 and 24, there is illustrated anotherembodiment of the web 170 shown in FIG. 17 used for the manufacture ofthe box 110, pouch 130 or bag 150 made in accordance with the presentinvention. In the embodiment shown an electro-photographic device 380employs magnetic brush technology to apply toner particles 400 comprisedof the metal-ion sequestering agents in a predetermined pattern 410 tothe web 170 forming a metal-ion sequestering web 350. The predeterminedpattern may comprise the entire surface of said web 170 or selectedlocations on said web 170. In the example shown the electro-photographicdevice 380 uses a photoconductor belt 405, which is driven in thedirection indicated by the arrow 495 by drive roller assembly 485. Thephotoconductor belt 405 is charged with a high voltage corona via acharger 420 and selectively discharged with light from a light source430 such as an LED array to form an electrostatic pattern 440. Theelectrostatic pattern 440 is developed with the negatively charged tonerparticles 435 by a magnetic brush 460 composed of the negatively chargedtoner particles 435 and magnetic carrier particles (not shown) on amagnetic roller 470. The charged toner particles 435 are thentransferred from the photoconductor belt 405 to the web 170 as a powderpattern 475 that is then fused to the web 170 by heated rollers 490 toform a fused pattern 480 as the web 170 moves in the direction of thearrow 495. By laying down a pattern the edges 500 shown in FIG. 24 maybe left clean to facilitate sealing of the box 110, pouch 130 or bag150. After the charged toner particles 435 have been transferred, thephotoconductor belt 405 is cleaned of any residual toner particles by acleaning station 505. As in the case of an electrophotographic printerthe amount or density of the toner particles transferred to the web canbe controlled by the amount of charge placed on the web. Using thistechnique, which is understood by those skilled in the art, the amountor density of metal-ion sequestering agents in the form of tonerparticles can likewise be controlled. This allows for providing themetal-ion sequestering agent at a designated location and at a desireddensity on web 350. Thus, the web 350 may be designed to meet thevarious needs of the user. For example, if more that one designatedmetal-ion is to be treated, more than one type metal-ion sequesteringagent can be applied to web 350. In addition the density may be variedto adjust to the amount to which the designated metal ion is present inthe liquid nutrient allowing for efficient use of both metal ionsequestering agents. In addition to providing different type metal-ionsequestering agents other type agents may be applied in combination, forexample, the electro-photographic device 350 may employ magnetic brushtechnology to apply toner particles 400 comprised of a metal-ionsequestering agent and an antimicrobial agent, as previously described,in a pattern 410 on to the web 170. The web 170 may be made of anysuitable material and of any suitable construction. For example but notby way of limitation, web 170 may be made of nylon, polyester, naturalor synthetic fabric material, plastic, and may be continuous, or of afabric type weave of one or more material fibers. It is important thatthe material allow for the adhering of the metal-ion sequestering agentand allow appropriate interaction with the liquid agent.

Referring now the FIGS. 25 and 26, there is shown a schematic of anapparatus made in accordance with the present invention for treating aliquid. The apparatus includes a holding tank 600 for holding the liquidnutrient 10 comprising an inlet port 605 with a valve 610, an outletport 615 with a valve 620, a web rack assembly 625 capable oftransporting the metal-ion sequestering web 350, as previouslydescribed, through the tank 602, a web supply 630, a web take up 635each having a web drive 640, such as motor (not shown) for transportingof the web 170 through the tank about web rack assembly 625. In theembodiment illustrated in FIG. 25 a fresh section of the metal-ionsequestering web 350 is transported from the web supply 630 as indicatedby arrows 645 into the empty holding tank 600 via the web rack assembly625 and web drive 630. Preferably the rack assembly provided aserpentine path along which the web travels allowing for a predeterminedamount of web to be subjected to the liquid for a predetermined amountof time. The holding tank 600 is then filled with an untreated batch ofa liquid nutrient 10 via the inlet port 615 and valve 610 as indicatedby arrow 650. The liquid nutrient 10 remains in the holding tank 600 andmay be recirculated through the tank 600 by a pump 647 until the amountof the specified metal ion(s) is substantially reduced to the desiredlevel as described in commonly assigned U.S. patent application Ser. No.10/985,393 filed concurrently herewith entitled CONTAINER FOR INHIBITINGMICROBIAL GROWTH IN LIQUID NUTRIENTS by David L. Patton et al. Forexample, but not limited to the designated metal be removed from theliquid nutrient 10 is reduced greater than 50%, preferably greater than80%. The time it takes to reduce the designated metal ion from theliquid nutrient will depend on a variety of factors, for example, thestarting amount of the designated metal ion in the nutrient, the rate ofrecirculation of the liquid nutrient by the web 30, the amount anddensity of metal-ion sequestering agent on the web 350, etc. Examples ofthe time for removing a designated metal-ion may take from one to 30minutes. The amount of time the liquid nutrient 10 resides in theholding tank 600 containing a treatment bed as described in commonlyassigned U.S. patent application Ser. No. 10/985,393 filed herewithentitled CONTAINER FOR INHIBITING MICROBIAL GROWTH IN LIQUID NUTRIENTSby David L. Patton et al. as previously discussed. After the desiredmetal ion level is reached, the liquid nutrient 10 is drained from theholding tank 600 via outlet port 615 and valve 620 as indicated by arrow652. After the tank 600 has been emptied, a new unused section of themetal-ion sequestering web 350 is transported into the tank 600 from theweb supply 630 and the used portion of metal-ion sequestering web 350 iswound onto the web take up 635 by the web drive 640. As the web 350exits the tank 600, a pair of squeegee rollers 670 remove any excessliquid nutrient 10 from the web 350. In yet another embodiment shown inFIG. 27, the liquid nutrient 10 is pumped into the holding tank 602,like numerals indicating like elements and operation as previouslydiscussed, through valve 610 and inlet port 605. In this embodiment theweb 170 moves along rack assembly 625 at a predetermined rate such thatthe web 170 will be treated to the appropriate level as it leaves tank600. The liquid nutrient 10 flows through the tank 600 as directed bytank baffles 660, the flow pattern of the liquid nutrient 10 beingindicated by arrows 665, until it exits out the outlet port 615 asindicated by arrow 652. The liquid nutrient's 10 flow is counter flow,opposite direction, with the path of the metal-ion sequestering web 350through the web rack assembly 625 as indicated by the arrows 667. As theweb 350 exists the tank 600 a pair of squeegee rollers 670 remove anyexcess liquid nutrient 10 from the web 350. It should be noted that themetal-ion sequestering web 350 may also be transported in the samedirection that the liquid nutrient 10 flow.

In another modified embodiment of the tank 600 and web transportassembly made in accordance with the present invention as illustrated inFIG. 28, like parts indicating like parts and operation as previouslydescribed. A sparger assembly 675 connected to the pump 647 impinges theliquid nutrient 10 against the metal-ion sequestering web 350 vianozzles 680 as the liquid nutrient 10 is recirculated back into tank602. The sparger assembly 675 may be used in any of the previouslydiscussed embodiments.

The invention has been described in detail with particular reference tocertain preferred embodiments thereof but it will be understood thatvariations and modifications can be effected within the spirit and scopeof the invention.

PARTS LIST

-   5 fluid container/bottle-   10 liquid nutrient-   15 inner polymeric layer-   20 outer polymeric layer-   22 barrier layer-   25 micro-organism-   30 “free” iron ion-   35 metal-ion sequestering agents-   35′ metal-ion sequestering agent with a sequestered metal ion-   40 hydrophilic polymer-   45 inner portion-   50 bottle cap-   52 insert-   55 hydrophilic polymer-   60 extension (straw)-   65 hydrophilic polymer-   80 inside surface-   85 supply tube-   90 spherical shaped nozzle assembly-   95 arrow-   100 arrow-   105 spray coating-   110 juice box-   115 inner layer-   120 middle layer-   125 outer layer-   130 pouch-   135 inner layer-   140 outer layer-   145 coating-   150 bag-   155 aqueous material-   160 inner layer-   165 outer layer-   170 base web-   175 hydrophilic layer-   180 coating assembly-   185 reservoir-   190 applicator-   200 can-   205 lining-   210 strip-   220 filter assembly-   225 inlet port-   230 outlet port-   235 filter-   240 arrow-   250 fluid bed ion exchange assembly-   255 holding tank-   260 inlet port-   265 outlet port-   270 fluid bed-   275 sequestering material-   280 solution-   285 arrow-   290 arrow-   295 arrow-   300 core material-   305 shell material-   310 “free” metal ions-   315 arrows-   320 gathered metal ions-   350 metal-ion sequestering web-   380 electro-photographic device-   400 toner particles-   405 photoconductor belt-   410 pattern-   415 arrow-   420 charger-   430 light source-   435 charged toner particles-   440 electrostatic pattern-   460 magnetic brush-   470 magnetic roller-   475 powder pattern-   480 fused pattern-   485 drive roller assembly-   490 heated rollers-   495 arrow-   500 edges-   505 cleaning station-   600 tank-   605 inlet port-   610 valve-   615 outlet port-   620 valve-   625 web rack assembly-   630 web supply-   635 web take up-   640 web drive-   645 arrow-   647 pump-   650 arrow-   652 arrow-   655 arrow-   660 tank baffles-   670 squeegee rollers-   675 nozzle

1. A method comprising the steps of: a. providing a tank for containinga liquid to be treated; b. providing a web in said tank, said web havingat least one metal-ion sequestering agent and/or antimicrobial agent; c.subjecting a liquid provided in said tank to said web; and d.recirculating said liquid through said tank, wherein said at least onemetal-ion sequestering agent and/or antimicrobial agent comprisesderivatized nanoparticles comprising inorganic nanoparticles having anattached metal-ion sequestrant, wherein said inorganic nanoparticleshave an average particle size of less than 200 nm and the derivatizednanoparticles have a stability constant greater than 10¹⁰ with iron(III).
 2. The method of claim 1 wherein said at least one metal-ionsequestering agent and/or antimicrobial agent is attached to thenanoparticle by reacting the nanoparticle with a silicon alkoxideintermediate of the sequestrant having the general formula:Si(OR)_(4-x)R′_(x;) wherein x is an integer from 1 to 3; R is an alkylgroup; and R′ is an organic group containing an alpha amino carboxylate,a hydroxamate, or a catechol.
 3. A method comprising the steps of: a.providing a tank for containing a liquid to be treated; b. providing aweb in said tank, said web having at least one metal-ion sequesteringagent and/or antimicrobial agent; and c. subjecting the liquid providedin said tank to said web, wherein said at least one metal-ionsequestering agent and/or antimicrobial agent comprises derivatizednanoparticles comprising inorganic nanoparticles having an attachedmetal-ion sequestrant, wherein said inorganic nanoparticles have anaverage particle size of less than 200 nm and the derivatizednanoparticles have a stability constant greater tan 10¹⁰ with iron(III).
 4. The method of claim 3 further comprising the step oftransporting said web along a rack provided in said tank at apredetermined rate.
 5. The method of claim 3 wherein said at least onemetal-ion sequestering agent and/or antimicrobial agent comprises atleast one metal-ion sequestering agent and at least one antimicrobialagent each provided at a different designated location on said web. 6.The method of claim 3 wherein said web is made of any one or more of thefollowing materials: plastic; nylon; polyester; and natural or syntheticfabric material.
 7. The method of claim 3 wherein said web comprises afabric weave made of one or more fiber materials.
 8. The method of claim3 further comprising the steps of: electrostatically charging a belt atdesignated locations so that said at least one metal-ion sequesteringagent and/or antimicrobial agent is magnetically charged so as to beattracted to the belt at said designated locations; and transferringsaid at least one metal-ion sequestering agent and/or antimicrobialagent from the belt to the web at predetermined locations on the webcorresponding to said designated locations.
 9. The method of claim 8wherein said designated locations comprise a predetermined pattern. 10.The method of claim 9 wherein said predetermined pattern comprisessubstantially the entire surface of said web.
 11. The method of claim 3wherein said at least one metal-ion sequestering agent and/orantimicrobial agent comprises at least two different metal-ionsequestering agents.
 12. The method of claim 3 wherein said at least onemetal-ion sequestering agent and/or antimicrobial agent comprises anantimicrobial agent and a metal-ion sequestering agent.
 13. The methodof claim 3 wherein said at least one metal-ion sequestering agent and/orantimicrobial agent is immobilized on at least one surface of said weband has a stability constant greater than 10¹⁰ with iron (III).
 14. Themethod of claim 3 wherein said at least one metal-ion sequestering agentand/or antimicrobial agent comprises an antimicrobial active materialselected from benzoic acid, sorbic acid, nisin, thymol, allicin,peroxides, imazalil, triclosan, benomyl, metal-ion release agents, metalcolloids, anhydrides, and organic quaternary animonium salts, a metalion exchange reagent including silver sodium zirconium phosphate, silverzeolite, or silver ion exchange resin.
 15. The method of claim 3 whereinsaid at least one metal-ion sequestering agent and/or antimicrobialagent comprises a metal ion selected from one of the following: silver;copper; gold; nickel; tin; and zinc.
 16. The method of claim 3 whereinsaid at least one metal-ion sequestering agent and/or antimicrobialagent is immobilized on at least one surface of said web and has astability constant greater than 10¹⁰ with iron (III) and saidantimicrobial agent comprises an antimicrobial active material selectedfrom benzoic acid, sorbic acid, nisin, thymol, allicin, peroxides,imazalil, triclosan, benomyl, metal-ion release agents, metal colloids,anhydrides, and organic quaternary ammonium salts.
 17. The method ofclaim 3 wherein said at least one metal-ion sequestering agent and/orantimicrobial agent is immobilized on at least one surface of said weband has a stability constant greater than 10¹⁰ with iron (III) and saidantimicrobial agent comprises a metal ion selected from one of thefollowing: silver; copper; gold; nickel; tin; and zinc.
 18. The methodof claim 3 wherein said at least one metal-ion sequestering agent and/orantimicrobial agent is immobilized on at least one surface of said weband has a high-affinity for biologically important metal ions includingMn, Mg, Zn, Cu and Fe.
 19. The method of claim 3 wherein said at leastone metal-ion sequestering agent and/or antimicrobial agent isimmobilized on at least one surface of said web and has ahigh-selectivity for biologically important metal ions including Mn, Mg,Zn, Cu and Fe.
 20. The method of claim 3 wherein said at least onemetal-ion sequestering agent and/or antimicrobial agent has ahigh-selectively for certain metal ions but a low-affinity for at leastone other ion.
 21. The method of claim 20 wherein said certain metalions comprises Mn, Mg, Zn, Cu and Fe and said other at least one ioncomprises calcium.
 22. The method of claim 3 wherein said at least onemetal-ion sequestering agent and/or antimicrobial agent is attached tothe nanoparticle by reacting the nanoparticle with a silicon alkoxideintermediate of the sequestrant having the general formula:Si(OR)_(4-x)R′_(x;) wherein x is an integer from 1 to 3; R is an alkylgroup; and R′ is an organic group containing an alpha amino carboxylate,a hydroxamate, or a catechol.